1. Field of the Invention
This invention concerns a fluorescence analysis method that uses a dye that changes from being non-fluorescent to fluorescent upon binding with an antibody.
2. Related Background Art of the Invention
The analysis of biological components and various chemical substances (medical drugs, environmental pollutants, etc.) in a living body is extremely important for academic research and the diagnosis and treatment of illnesses. However, since generally-used prior-art methods for analyzing these substances require the sampling of body fluid, extraction of tissue, and complex, time-consuming preprocesses for removing foreign substances, the development of analysis methods that enable in vivo substances, etc., to be analyzed simply and yet at high sensitivity were desired.
Immunoassay (immunoassay and immunometric assay) methods using fluorescent dye labeled antibodies have thus been developed as one of the methods enabling such analysis. By such an immunoassay method, analysis of high selectivity is enabled by a specific molecule recognition function based on an antigen-antibody reaction and selective measurement of the analyte is enabled by fluorescence analysis without removing foreign substances. An antibody, which has been labeled in advance by a fluorescent dye, is generally used as the fluorescent dye labeled antibody, and a single substance (antigen) is recognized specifically by an antigen-binding site. In Japanese Laid-Open Patent Application No. 4-211363 is disclosed a bispecific hybrid monoclonal antibody having two antigen-binding sites, one of which is induced by a tissue plasminogen activator and the other of which is induced by a fluorescent substance or other label. In Japanese Laid-Open Patent Application No. 9-5324 is disclosed an antibody for an antigen formed by adding a non-fluorescent dye to an immunity substance. Furthermore, in Japanese Laid-Open Patent Application No. 11-183477 is disclosed a fluorescence immunoassay method, with which a fluorescence intensity change is measured under the presence of insulin, based on an antigen-antibody reaction between a covalently bound substance (MG-labeled insulin) of insulin and malachite green (MG) and an anti-MG-Ins antibody, for which this covalently bound substance is an antigen.